Review TRENDS in Biotechnology Vol.22 No.

10 October 2004

Glucose and sucrose: hazardous

fast-food for industrial yeast?
Kevin J. Verstrepen1,2, Dirk Iserentant3, Philippe Malcorps4, Guy Derdelinckx2,
Patrick Van Dijck5, Joris Winderickx6, Isak S. Pretorius7, Johan M. Thevelein5 and
Freddy R. Delvaux2
1
MIT Whitehead Institute for Biomedical Research, Nine Cambridge Center, Cambridge, MA 02142, USA
2
Centre for Malting and Brewing Science, Department of Food and Microbial Technology, Katholieke Universiteit Leuven,
Kasteelpark Arenberg 22, B-3001 Leuven (Heverlee), Belgium
3
Flanders Interuniversitary Institute for Biotechnology (V.I.B.), Rijvisschestraat 120, B-9052 Zwijnaarde, Belgium
4
Interbrew NV, Vaartstraat 94, B-3000 Leuven, Belgium
5
Laboratory of Molecular Cell Biology, Department of Biology, Katholieke Universiteit Leuven,
and Department of Molecular Microbiology, Flanders Interuniversitary Institute for Biotechnology (V.I.B.),
Kasteelpark Arenberg 31, B-3001 Leuven (Heverlee), Belgium
6
Functional Biology, Department of Biology, Katholieke Universiteit Leuven, Kasteelpark Arenberg 31, B-3001 Leuven (Heverlee),
Belgium
7
The Australian Wine Research Institute, PO Box 197, Glen Osmond, Adelaide SA-5064, Australia

Yeast cells often encounter a mixture of different biotechnology, including alcoholic fermentation, bread pro-
carbohydrates in industrial processes. However, glucose duction and the fabrication of biopharmaceuticals.
and sucrose are always consumed first. The presence of In this review, we begin by summarizing the role of the
these sugars causes repression of gluconeogenesis, the two best-known glucose-triggered signaling cascades in
glyoxylate cycle, respiration and the uptake of less- S. cerevisiae: the main glucose repression pathway (also
preferred carbohydrates. Glucose and sucrose also known as the catabolite repression pathway), and the
trigger unexpected, hormone-like effects, including the Ras/cAMP/protein kinase A (PKA) pathway. We then
activation of cellular growth, the mobilization of storage consider the role and implications of sugar signaling in
compounds and the diminution of cellular stress resist- industrial yeast-based processes.
ance. In an industrial context, these effects lead to
several yeast-related problems, such as slow or incom-
Glucose regulation of carbohydrate uptake and
plete fermentation, ‘off flavors’ and poor maintenance of
metabolism
yeast vitality. Recent studies indicate that the use of
In response to glucose and sucrose, the main glucose
mutants with altered responses to carbohydrates can
repression pathway downregulates several genes
significantly increase productivity. Alternatively, avoid-
involved in the uptake and metabolism of alternative
ing unnecessary exposure to glucose and sucrose could
carbohydrates, as well as genes involved in gluconeo-
also improve the performance of industrial yeasts.
genesis and respiration. The repression of respiration by
glucose and sucrose, which is known as the ‘Crabtree
Unlike most mammalian cells, which function in an effect’, might seem counterproductive but, although
environment with virtually constant concentrations of respiration is a more efficient method of energy
only one sugar, namely glucose, yeast cells encounter production, fermentation offers the advantage of ethanol
variable concentrations of many different carbon sources. production, which hampers the growth of competing
To overcome this lack of homeostasis and to survive in a microorganisms. In addition, glucose concentrations,
highly competitive ecosystem, the common brewer’s and and possibly levels of glycolytic intermediates, regulate
baker’s yeast Saccharomyces cerevisiae has evolved the the expression of several glucose transporters and some
capacity to take up and to metabolize different carbo- glycolytic genes (Figure 1 and reviewed in Refs [1–4]).
hydrates. S. cerevisiae cells also have several mechanisms Thus, the main glucose repression pathway ensures that
for sensing the nutritional status of the environment, the preferred sugars are metabolized before the con-
enabling them to adapt their uptake and metabolism of sumption of alternative carbohydrates, such as maltose
nutrients to specific conditions. Although these signaling and galactose.
pathways have been studied mostly as a model for nutrient- Glucose also slows down the uptake of fructose
induced signaling cascades in higher eukaryotes, their because both sugars are imported by the same carriers,
unraveling has relevance for several applications of yeast which have a greater affinity for glucose than for
fructose. Besides this competitive inhibition of fructose
Corresponding author: Kevin J. Verstrepen ([email protected]). uptake, recent research shows that glucose can repress
Available online 28 August 2004 the expression of specific fructose transporters such as
www.sciencedirect.com 0167-7799/$ - see front matter Q 2004 Elsevier Ltd. All rights reserved. doi:10.1016/j.tibtech.2004.08.001
532 Review TRENDS in Biotechnology Vol.22 No.10 October 2004

Figure 1. The main glucose repression pathway in Saccharomyces cerevisiae. Figure 2. The Ras/cAMP/protein kinase A (PKA) nutrient signaling pathway in
Extracellular glucose is taken up through one of the hexose transporters (Hxt) and Saccharomyces cerevisiae. Full activation of this pathway requires a dual signal.
subsequently phosphorylated to glucose 6-phosphate (glucose-6P) by one of the First, the intracellular phosphorylation of glucose to glucose 6-phopshate (glucose-
hexokinases (Hxk). This phosphorylation process and/or the depletion of AMP 6P) enhances the activity of adenylate cyclase Cyr1. Second, extracellular glucose
owing to the increased production of ATP inactivates the central protein kinase Snf1 or sucrose is sensed by a G-protein-coupled receptor system, consisting of the
[59]. Inactivation of Snf1 occurs via either inhibition of its phosphorylation or receptor Gpr1 and the Ga protein Gpa2 [10]. This second signal strongly enhances
stimulation of the Snf1-dephosphorylation activity of the phosphatase complex PP1 Cyr1 activity, resulting in a transient peak of cAMP immediately after the addition of
[60,61]. When Snf1 is inactive, the DNA-binding protein Mig1 is translocated from glucose or sucrose [9,10,57,64]. The rise in cAMP causes the regulatory subunits
the cytoplasm to the nucleus [62]. In the nucleus, Mig1 recruits the general repres- (Bcy1) of the PKA complex to bind cAMP and to dissociate from the Tpk catalytic
sors Tup1 and Ssn6 and binds to the promoters of several glucose-repressed genes, subunits of PKA [65,66]. The free Tpk kinases then phosphorylate various target
including genes involved in gluconeogenesis, respiration and the uptake and proteins, which eventually leads to acclimatization to high glucose levels (Figure 3).
breakdown of alternative carbon sources, such as maltose (MAL genes) and Although the role of the small G proteins Ras1 and Ras2 in this process is not
galactose (GAL genes). When extracellular glucose is depleted, the Snf1 complex completely understood, these proteins are essential for basal Cyr1 activity [67,68].
is activated, causing the translocation of Mig1 back to the cytoplasm, where it can For details, see reviews by Johnston [2] and Winderickx et al. [4]. For clarity, note
no longer repress its targets [62,63]. Thus, glucose repression is relieved and alter- that Gpa2 and Cyr1 are not depicted as attached to the plasma membrane and the
native carbon sources can be taken up. Fructose and sucrose, which is extracellularly shuttling of the free PKA catalytic subunits out of the nucleus [69] is not shown.
hydrolyzed into glucose and fructose, exert catabolite repression effects similar to
those of glucose, although glucose is taken up preferentially [1]. For details, see
reviews by Gancedo [1], Johnston [2], Carlson [3] and Winderickx et al. [4]. pathway requires extracellular glucose or sucrose [10],
other sugars such as fructose, maltose, maltotriose and
Fsy1 [5–7]. In addition to regulating the uptake of galactose cannot trigger a strong cAMP/PKA response
alternative sugars, the main glucose repression pathway [10,11] (Figure 2).
prevents futile cycling in carbohydrate metabolism by Among the targets regulated by the Ras/cAMP/PKA
shutting down de novo synthesis of glucose by the pathway are genes encoding heat-shock proteins, such as
gluconeogenic pathway. HSP12 and HSP104, which are rapidly repressed on
activation of this pathway [12,13] (Figure 3). These
proteins have important roles in various processes that
Regulation of the hormone-like effects of glucose and help yeast cells to cope with a broad array of stresses,
sucrose including heat and ethanol stress [14–16]. Furthermore,
Another principal carbon signaling pathway, the high PKA activity also causes repression of the TPS1 and
Ras/cAMP/PKA pathway, controls the expression of TPS2 genes encoding trehalose synthase [17]. Trehalose
various genes involved in metabolism, proliferation and has a prominent role in cellular stress resistance because
stress resistance in response to glucose (Figure 2 and it protects membranes from desiccation and prevents
reviewed in Refs [4,8,9]). Because full activation of this protein denaturation [18].
www.sciencedirect.com
 Review TRENDS in Biotechnology Vol.22 No.10 October 2004 533

The negative effects of catabolite repression on yeast

performance
In an industrial context, one of the best-known adverse
effects of the main glucose repression pathway on yeast
performance is seen in the production of baker’s yeast,
during which the switch from respiration to fermentation
induced by glucose or sucrose causes a marked drop in
biomass yield. Similarly, in the production of biopharma-
ceuticals, repression of respiration decreases the yield of
product [23,24]. Furthermore, the ethanol generated
during fermentation is an additional stress factor for the
yeast cells and can also negatively affect the stability of
the pharmaceutical product. To avoid these effects, yeasts
are grown mostly in aerated fed-batch reactors, in which
the glucose and sucrose concentrations are maintained
constantly below the threshold concentration for inducing
the Crabtree effect [23,25].
By contrast, the production of alcoholic beverages and
bioethanol requires fermentation. Apart from the first few
hours, these fermentations are carried out anaerobically
so that, even in the absence of the Crabtree effect, the
yeast cells cannot respire. In this set-up, the negative
effect of the main glucose repression pathway lies in the
repressed uptake and metabolism of other sugars, such as
galactose, maltose and maltotriose. The resulting reduced
consumption of alternative sugars negatively affects the
fermentation rate [26]. Remarkably, catabolite repression
is not always relieved immediately once glucose and
sucrose are depleted; instead, an initial preincubation in
glucose leads to the slower metabolism of other sugars
over several hours [21].
Figure 3. Targets of the Ras/cAMP/PKA nutrient signaling pathway. Targets include Empirical data obtained in various breweries show that
key proteins involved in the control of cell growth, glucose metabolism, stress
resistance (including trehalose metabolism), flocculation and filamentous growth,
extended propagation and cultivation in sucrose often
and volatile ester synthesis, as well as feedback mechanisms. For example, one leads to a permanent loss of maltose-fermenting capacity.
target of this pathway is the phosphodiesterase Pde1, which is responsible for the Similarly, it has been recently shown that several genes
rapid breakdown of cAMP [4].
encoding maltose transporters and maltase remain
repressed when yeast cells are inoculated into maltose-
containing medium after a long-term cultivation in
Glucose and sucrose signaling in industrial
glucose-rich medium [27]. This continued repression
fermentations
suggests that sustained favorable growth conditions,
In most commercial processes, the fermentation medium
such as the availability of glucose, might cause a long-
is a complex mixture of different fermentable sugars. The
term decrease in the capacity of yeast to metabolize
main sugars in grape must are glucose and fructose; by
alternative carbon sources.
contrast, beer wort contains glucose, fructose, sucrose,
maltose and maltotriose, and the fermentation medium
The negative effects of Ras/cAMP/PKA activation on
for bioethanol production is usually a mixture of any of
yeast performance
these sugars in variable concentrations, depending on the As mentioned above, in addition to catabolite repres-
origin of the molasses [19,20]. sion, glucose and sucrose also trigger activation of the
In most applications, the initial concentration of Ras/cAMP/PKA pathway, which in turn leads to a
glucose and/or sucrose in the growth medium is above decrease in the stress resistance of yeast cells. During
the threshold concentrations (20–40 mM) for inducing the industrial processes, yeast cells encounter various severe
sugar signaling cascades [21,22]. Thus, both the main stress conditions, including shear stress; marked shifts
glucose repression pathway and the Ras/cAMP/PKA in oxygen levels, temperature, osmolarity and pH; high
pathway are triggered at the start of the process. ethanol and carbon dioxide concentrations; and a lack of
Activation of these pathways has three main conse- nutrients [28,29]. Notably, Brosnan et al. [30] and
quences: repression of respiration, arrest of the con- Rossignol et al. [31] have reported that industrial yeast
sumption of other carbohydrates, and loss of cellular strains do not fully activate several stress responses
stress resistance. Although these effects presumably help during the first phases of industrial fermentations, which
S. cerevisiae to survive in its natural habitat, sugar could be due to the presence of glucose and sucrose.
signaling causes several problems in various yeast-based The inability of yeast cells to respond to unfavorable,
industrial processes. stressful conditions leads to sluggish fermentations and
www.sciencedirect.com
534 Review TRENDS in Biotechnology Vol.22 No.10 October 2004

cell autolysis [32]. The latter process causes severe competed the ancestral parent strain over long periods of
off flavors in alcoholic beverages, including a highly time under stressful industrial conditions [46]. Although
unpleasant goat-like aroma caused by the release of fatty this natural selection can lead to vast improvements in
acids, and a decrease in fruity aromas owing to the yeast performance, especially when yeasts are re-used for
hydrolysis of volatile esters by esterases leaking out of multiple fermentations, as they are in beer production,
damaged cells [33,34]. Furthermore, sustained high further improvements can be achieved by a more direct
concentrations of glucose cause a decrease in the replica- approach; for example, the use of mutagenesis by ultra-
tive lifespan of yeast, whereas the use of maltose-rich violet irradiation or chemical mutagens such as ethyl
media generally leads to an increase in cell longevity methane sulfonate, followed by a screening or selection
[35,36]. Other reported effects of high glucose include an procedure. Alternatively, superior yeast strains can be
increase in the instability of short chromosomes [37] and a developed by genetic engineering. Both methods have
strong increase in the production of aroma-active esters their pros and cons, which have been extensively dis-
through activation of the Ras/cAMP/PKA pathway [38,39]. cussed elsewhere [47,48]. Mutagenesis followed by selec-
Although moderate levels of these volatile esters are tion does not require any knowledge of target genes or
essential for the desired fruity flavors of beer and wine, metabolic pathways and is relatively easy to perform.
high glucose concentrations lead to an undesirable over- Furthermore, strains acquired through this process are
production of these compounds [39]. not considered as ‘genetically modified’, which makes
In addition to the Ras/cAMP/PKA, other closely related them virtually ready to use; however, a good selection
nutrient signaling pathways, such as the ‘target of procedure is required to facilitate rapid isolation of the few
rapamycin’ pathway and the ‘fermentable growth med- useful mutants from billions of ‘uninteresting’ cells. This
ium-induced’ pathway can influence the fermentation selection requirement severely limits the use of this
performance of yeast. In contrast to the short-term method for the isolation of yeasts with specific fermenta-
response of the Ras/cAMP/PKA pathway, which essen- tion characteristics.
tially responds to changes in glucose and sucrose levels, Many of the stress responses are not specific for
these pathways are thought to integrate the overall particular stress conditions [13,16]; thus, a potential
nutritional and/or energy status of cells [4,40,41]. This strategy is the selection of mutants that are more resistant
implies that, even in the presence of glucose and sucrose, to lethal stresses, such as heat or ethanol stress, in the
some cellular stress responses can be activated when other presence of glucose or sucrose. After this first selection,
essential nutrients (e.g. nitrogen sources) are depleted, as the isolated mutants can be screened for improved
can occur in the last phase of wine fermentations [42]. fermentation properties under industrial stress con-
Given the limited data, however, it is premature to esti- ditions. In our experience, approaches in which the actual
mate the importance of these signaling cascades in stressful fermentation conditions are mimicked as closely
industrial processes. as possible greatly increase the chance of success [49].
Ideally, all screens should be therefore done in pilot-scale
Yeast carbon signaling mutants with improved fermenters under growth conditions and medium similar
industrial performance to those used in the industrial-scale process. Even so, the
Considering the various adverse effects of carbohydrate mutant that performs best in pilot-scale trials is often not
signaling on the performance of industrial yeasts, the use the best strain for full-scale production. Thus, it is vital to
of strains with altered responses to glucose and sucrose select several mutants for production-scale trials.
should lead to a significant improvement in productivity. Recent progress in our understanding of the yeast
In addition, mutations in the regulatory systems might carbohydrate signaling pathways makes it possible to
provoke ‘balanced’ changes in the expression of several improve strains by genetic engineering, leading to the
genes involved in fermentation performance [43,44]. introduction of such traits as increased trehalose levels
Indirect phenotypic evidence suggests that many of the and constitutive expression of targeted carbohydrate
currently used industrial strains have already acquired transporters [48,50,51]. Brewers and wine producers are
altered sugar signaling properties. Some brewer’s strains, reluctant to use genetically modified yeasts, however,
for example, do not break down trehalose in the presence mainly because of the complex legislation and the negative
of glucose [45]. Furthermore, many brewer’s strains public perception. Furthermore, drastic changes in the
show constitutive uptake of maltose, independent of the vital carbon signaling pathways frequently affect other
presence of glucose [21]. Interestingly, in most cases commercially important properties.
glucose-mediated repression of the uptake of other less- The mutations found in superior strains selected after
preferred sugars, such as galactose, is still present. This random mutagenesis are often more subtle and can lead to
indicates that if the alternative sugar-usage patterns are more balanced performances [49,52]. Moreover, tech-
caused by alterations in the glucose-sensing pathways, niques similar to genome shuffling in bacteria [53] provide
these alterations are most probably present in a down- an efficient way with which to combine different subtle
stream part of the pathway [21]. Taken together, many improvements. Thus, although genetic engineering and
industrial strains have acquired alternative glucose self-cloning are certainly the most promising methods for
signaling phenotypes, enabling them to complete their the future, random mutagenesis and selection remain the
tasks better in industrial processes. most realistic method with which to obtain improved
Most of these yeasts are not the result of a specific, yeast strains for short-term introduction into industrial
intentional selection procedure, but have presumably out- food production.
www.sciencedirect.com
 Review TRENDS in Biotechnology Vol.22 No.10 October 2004 535

These arguments do not apply to the production of A healthy diet for optimal performance?
heterologous proteins and biopharmaceuticals, in which The selection or creation of superior yeast strains is not
the use of genetically modified organisms is less con- the only way to improve the productivity of industrial
troversial. Moreover, the growth medium often contains yeasts. Relatively simple changes in yeast handling can
only a single carbon source in these processes, and thus also lead to significant improvements. Considering the
the metabolic and regulatory complications associated many adverse effects of glucose and sucrose on yeast
with complex sugar mixtures are avoided. A good example, performance, it is advisable to avoid using high con-
developed and patented by Boles et al. [54], is a strain that centrations of these sugars. In many applications, the
has been genetically modified to express a chimeric hexose composition of the fermentation medium cannot be
transporter. This strain is relieved of the Crabtree effect, changed without affecting product quality and cost, but
and thus there is less need for sophisticated cultivation sometimes it is possible to use alternative carbon sources.
techniques to reach optimal production rates. Another For example, when high-carbon syrups are added to the
interesting strain has been developed by Ostergaard et al. medium, as is often done in high-gravity beer fermenta-
[43], who have used genetic engineering of the GAL tion or the production of bioethanol or biopharmaceuti-
regulatory system to increase galactose metabolism. cals, it might be better to use maltose-rich syrups instead
of sucrose-containing syrups.
Furthermore, when yeasts are propagated for indus-
Selecting interesting carbon signaling mutants: a trial use, it is beneficial to combine glucose or sucrose with
practical example alternative sugars, such as maltose syrups, or with non-
Relatively few carbon signaling mutants of industrial fermentable carbon sources, such as mannitol or sorbitol
yeast strains have been described or investigated, in part, [58]. The preferred sugars will be used first, facilitating
owing to the polyploid or aneuploid nature of these maximal biomass production rates, whereas the maltose
strains, which makes their genetic analysis much more or mannitol will be metabolized just before the production
difficult as compared with haploid laboratory strains. phase or storage of the yeast cells. Avoiding contact with
Perhaps the best examples of carbon signaling mutants glucose and sucrose just before the fermentation process
are the so-called ‘fermentation-induced loss of stress might lead to increased levels of stored carbohydrates,
resistance’ (fil) mutants of industrial baker’s yeast, better stress resistance and a faster fermentation rate. In
which show improved resistance to freezing and thawing the specific case of bioethanol production, it has proved
in the presence of glucose and sucrose. advantageous to remove sugar monomers continuously
Freeze–thaw resistance is required for the increasingly from the saccharification vessel to maximize the break-
popular use of frozen dough, which permits the separation down and use of more complex sugars [46].
of dough production and baking. Before freezing, yeast is
mixed with the dough. Because of the high glucose and/or Conclusion
sucrose concentrations in the dough, however, yeast cells In the constant strive for better and more efficient yeast-
rapidly activate the Ras/cAMP/PKA pathway and lose based industrial processes, much attention has been
their stress resistance, resulting in considerable loss of drawn to various high-tech solutions ranging from
vitality during freezing and thawing [49,55]. By irradiat- optimization of bioreactors to the refinement of genetic
ing baker’s yeast with ultraviolet, preparing dough with technologies. The recently unraveled negative effects of
the mutants and subsequently subjecting the dough to glucose and sucrose on the industrial performance of yeast
numerous freeze–thaw cycles, it has been possible to are often overlooked, and these two sugars remain among
isolate several mutants with improved freeze tolerance, the most commonly used carbon sources in yeast media.
although only a few strains retain all of the other desirable In general, the presence of these sugars at moderate
characteristics of industrial baker’s yeast and perform concentrations is inevitable and will not cause significant
superiorly in full-scale processes of dough freezing and problems. In some applications, however, high concen-
thawing [49,56]. The exact mutations in these industrial trations of glucose or sucrose at a particular stage of the
strains are not known. industrial process cause a significant and long-term
fil mutants have been also isolated from laboratory decrease in fermentation performance. Thus, the use of
yeast strains, and two of these mutations have been sugar signaling mutants or the reduction of glucose and
identified. One strain contains a mutation in the GPR1 sucrose might be advantageous in various bioindustries,
gene, which encodes a G-protein-coupled receptor ranging from the production of alcoholic beverages and
involved in glucose and sucrose sensing [57] (Figure 2). bread to the fabrication of biopharmaceuticals. Indeed,
The other strain contains a partially inactivating although yeast cells seem to prefer glucose and sucrose
mutation in the CYR1 gene encoding adenylate cyclase, over other carbohydrates, offering them too much of this
which is also an essential component of the Ras/cAMP/ ‘fast-food’ negatively affects their general fitness, again
PKA pathway [44,52]. These mutations in Ras/cAMP/PKA demonstrating the striking analogy between yeasts and
signaling prevent the strains from losing their stress higher eukaryotes.
resistance in glucose and sucrose media. Remarkably,
some of the laboratory fil strains were selected for
Acknowledgements
heat-resistance in glucose medium, but the superior We thank Tom DiCesare for assistance with graphics, and An Jansen,
mutants also show improved resistance to various other Reeta Prusty and Sherwin Chan for discussions. Kevin J. Verstrepen is a
stresses [44,52]. Research Assistant of the Fund for Scientific Research, Flanders
www.sciencedirect.com
536 Review TRENDS in Biotechnology Vol.22 No.10 October 2004

(Belgium) (FWO-Vlaanderen), a Hoover Foundation Fellow and a 22 Meijer-Michelle, M.C. et al. (1998) Glucose repression in Saccharo-
D. Collen Fellow of the Belgian American Educational Foundation. myces cerevisiae is related to the glucose concentration rather than the
Original work in our laboratories was supported by grants from the Fund glucose flux. J. Biol. Chem. 273, 24102–24107
for Scientific Research - Flanders (FWO project G.0082.03), the Research 23 Calado, C.R.C. et al. (2003) Development of a fed-batch cultivation
Fund of the K.U. Leuven (O.T. project 03/40, Concerted Research Actions strategy for the enhanced production and secretion of cutinase by
GOA 2002/05 and Flanders–South Africa project BIL02/34) and Inter- recombinant Saccharomyces cerevisiae SU50 strain. J. Biosci. Bioeng.
university Attraction Poles Network P5/30. 96, 141–148
24 Ejiofor, A.O. et al. (1994) A robust fed-batch feeding strategy for
optimal parameter estimation for baker’s yeast production. Bioprocess
Eng. 11, 135–144
References
25 Valentinotti, S. et al. (2003) Optimal operation of fed-batch fermenta-
1 Gancedo, J.M. (1998) Yeast carbon catabolite repression. Microbiol.
tions via adaptive control of overflow metabolite. Control Eng. Pract.
Mol. Biol. Rev. 62, 334–361
11, 665–674
2 Johnston, M. (1999) Feasting, fasting and fermenting – glucose
26 Shimizu, H. et al. (2002) Effect of carbon and nitrogen additions on
sensing in yeast and other cells. Trends Genet. 15, 29–33
consumption activity of apparent extract of yeast cells in a brewing
3 Carlson, M. (1999) Glucose repression in yeast. Curr. Opin. Microbiol.
process. J. Am. Soc. Brew. Chem. 60, 163–169
2, 202–207
27 Kuthan, M. et al. (2003) Domestication of wild Saccharomyces
4 Winderickx, J. et al. (2003) From feast to famine: adaptation to
cerevisiae is accompanied by changes in gene expression and colony
nutrient availability in yeast. In Topics in Current Genetics, Vol. 1:
morphology. Mol. Microbiol. 47, 745–754
Yeast Stress Responses (Hohmann, S. and Mager, P.W.H., eds),
28 Attfield, P.V. (1997) Stress tolerance: the key to effective strains of
pp. 305–386, Springer-Verlag
industrial baker’s yeast. Nat. Biotechnol. 15, 1351–1357
5 Berthels, N.J. et al. (2004) Discrepancy in glucose and fructose
29 Bauer, F.F. and Pretorius, I.S. (2000) Yeast stress response and
utilisation during fermentation by Saccharomyces cerevisiae wine
fermentation efficiency: how to survive the making of wine – a review.
yeast strains. FEMS Yeast Res. 4, 683–689
S. Afr. J. Enol. Vitic. 21, 27–51
6 de Sousa, H.R. et al. (2004) Differential regulation by glucose and
30 Brosnan, M.P. et al. (2000) The stress response is repressed during
fructose of a gene encoding a specific fructose/HC symporter in
fermentation of brewery strains of yeast. J. Appl. Microbiol. 88,
Saccharomyces cerevisiae. Yeast 21, 519–530
746–755
7 Goncalves, P. et al. (2000) FSY1, a novel gene encoding a specific
31 Rossignol, T. et al. (2003) Genome-wide monitoring of wine yeast gene
fructose/HC symporter in the type strain of Saccharomyces carlsber-
expression during alcoholic fermentation. Yeast 20, 1369–1385
gensis. J. Bacteriol. 182, 5628–5630
8 Thevelein, J.M. and De Winde, J.H. (1999) Novel sensing mechanisms 32 Ivorra, C. et al. (1999) An inverse correlation between stress
and targets for the cAMP-protein kinase A pathway in the yeast resistance and stuck fermentations in yeast. A molecular study.
Saccharomyces cerevisiae. Mol. Microbiol. 33, 904–918 Biotechnol. Bioeng. 64, 698–708
9 Versele, M. et al. (2001) Sex and sugar in yeast: two distinct GPCR 33 Neven, H. et al. (1997) Flavor evolution of top fermented beers. MBAA
systems. EMBO Rep. 2, 574–579 Tech. Quart. 34, 115–118
10 Rolland, F. et al. (2000) Glucose-induced cAMP signalling in yeast 34 Taylor, G.T. and Kirsop, B.H. (1977) The origin of medium chain
requires both a G-protein coupled receptor system for extracellular length fatty acids present in beer. J. Inst. Brew. 83, 241–243
glucose detection and a separable hexose kinase-dependent sensing 35 Maskell, D.L. et al. (2001) Impact of carbohydrate composition of
process. Mol. Microbiol. 38, 348–358 media on lager yeast replicative lifespan. J. Am. Soc. Brew. Chem. 59,
11 Rolland, F. et al. (2001) The role of hexose transport and phosphoryl- 111–116
ation in cAMP signalling in the yeast Saccharomyces cerevisiae. 36 Lin, S.J. et al. (2002) Calorie restriction extends Saccharomyces
FEMS Yeast Res. 1, 34–45 cerevisiae lifespan by increasing respiration. Nature 418, 344–348
12 Varela, J.C.S. et al. (1995) The Saccharomyces cerevisiae HSP12 gene 37 Sato, M. et al. (2002) Effect of growth media and strains on structural
is activated by the high-osmolarity glycerol pathway and negatively stability in small chromosomes (chromosomes I, VI and III) of bottom
regulated by protein kinase A. Mol. Cell. Biol. 15, 6232–6245 fermenting yeast. J. Inst. Brew. 108, 283–285
13 Marchler, G. et al. (1993) A Saccharomyces cerevisiae UAS element 38 Verstrepen, K.J. et al. (2003) Flavour-active esters: adding fruitiness
controlled by protein kinase A activates transcription in response to a to beer. J. Biosci. Bioeng. 96, 110–118
variety of stress conditions. EMBO J. 12, 1997–2003 39 Verstrepen, K.J. et al. (2003) The Saccharomyces cerevisiae alcohol
14 Piper, P.W. et al. (1997) Hsp30, the integral plasma membrane heat acetyl transferase gene ATF1 is a target of the cAMP/PKA and FGM
shock protein of Saccharomyces cerevisiae, is a stress-inducible nutrient signalling pathways. FEMS Yeast Res. 4, 285–296
regulator of plasma membrane HC-ATPase. Cell Stress Chaperones 40 Crauwels, M. et al. (1997) The Sch9 protein kinase in the yeast
2, 12–24 Saccharomyces cerevisiae controls cAPK activity and is required for
15 Sanchez, Y. et al. (1992) HSP104 is required for tolerance to many activation of the fermentable-growth-medium-induced (FGM) path-
forms of stress. EMBO J. 11, 2357–2364 way. Microbiol. 143, 2627–2637
16 Piper, P.W. (1995) The heat-shock and ethanol stress responses of 41 Pedruzzi, I. et al. (2003) TOR and PKA signalling pathways con-
yeast exhibit extensive similarity and functional overlap. FEMS verge on the protein kinase Rim15 to control entry into G0. Mol.
Microbiol. Lett. 134, 121–127 Cell 12, 1–20
17 Winderickx, J. et al. (1996) Regulation of genes encoding subunits of 42 Puig, S. and Pérez-Ortin, J.E. (2000) Stress response and expression
the trehalose synthase complex in Saccharomyces cerevisiae: novel patterns in wine fermentations of yeast genes induced at the diauxic
variations of STRE-mediated transcription control? Mol. Gen. Genet. shift. Yeast 16, 139–148
252, 470–482 43 Ostergaard, S. et al. (2000) Increasing galactose consumption by
18 Wiemken, A. (1990) Trehalose in yeast: stress protectant rather than Saccharomyces cerevisiae through metabolic engineering of the GAL
reserve carbohydrate. Antonie Van Leeuwenhoek Int. J. Gen. Mol. gene regulatory network. Nat. Biotechnol. 18, 1283–1286
Microbiol. 58, 209–217 44 Versele, M. et al. (2004) The high general stress resistance of the
19 Yoon, S-H. et al. (2003) Specificity of yeast (Saccharomyces cerevisiae) Saccharomyces cerevisiae fil1 adenylate cyclase mutant (Cyr1Lys1682)
in removing carbohydrates by fermentation. Carbohydr. Res. 338, is only partially dependent on trehalose, Hsp104 and overexpression
1127–1132 of Msn2/4-regulated genes. Yeast 21, 75–86
20 Bamforth, C.W. (2003) Wort composition and beer quality. In Brewing 45 Reinman, M. and Londesborough, J. (2000) Rapid mobilization of
Yeast Fermentation Performance (Vol. 2) (Smart, K. ed.), pp. 77–85, intracellular trehalose by fermentable sugars: a comparison of
Blackwell Science different strains. In Brewing Yeast Fermentation Performance
21 Meneses, J.F. et al. (2002) A survey of industrial strains of (Vol. 1, 1st edn) (Smart, K. ed.), Blackwell Science
Saccharomyces cerevisiae reveals numerous altered patterns of 46 Wheals, A.E. et al. (1999) Fuel ethanol after 25 years. Trends
maltose and sucrose utilisation. J. Inst. Brew. 108, 310–321 Biotechnol. 17, 482–487
www.sciencedirect.com
 Review TRENDS in Biotechnology Vol.22 No.10 October 2004 537

47 Pretorius, I.S. (2000) Tailoring wine yeast for the new millennium: 58 Quain, D.E. and Boulton, B. (1987) Growth and metabolism of
novel approaches to the ancient art of winemaking. Yeast 16, 675–729 mannitol by strains of Saccharomyces cerevisiae. J. Gen. Microbiol.
48 Dequin, S. (2001) The potential of genetic engineering for improving 133, 1675–1684
brewing, wine-making and baking yeasts. Appl. Microbiol. Biotechnol. 59 Wilson, W.A. et al. (1996) Glucose repression/derepression in budding
56, 577–588 yeast: SNF1 protein kinase is activated by phosphorylation under
49 Teunissen, A. et al. (2002) Isolation and characterization of a freeze- derepressing conditions, and this correlates with a high AMP:ATP
tolerant diploid derivative of an industrial baker’s yeast strain and its ratio. Curr. Biol. 6, 1426–1434
use in frozen doughs. Appl. Environ. Microbiol. 68, 4780–4787 60 Ludin, K. et al. (1998) Glucose-regulated interaction of a regulatory
50 Pretorius, I.S. and Bauer, F.F. (2002) Meeting the consumer challenge subunit of protein phosphatase 1 with the Snf1 protein kinase in
through genetically customized wine-yeast strains. Trends Biotechnol. Saccharomyces cerevisiae. Proc. Natl. Acad. Sci. U. S. A. 95, 6245–6250
20, 426–432 61 Sanz, P. et al. (2000) Sip5 interacts with both the Reg1/Glc7 protein
51 Verstrepen, K.J. et al. (2001) Genetic modification of Saccharomyces phosphatase and the Snf1 protein kinase of Saccharomyces cerevisiae.
cerevisiae: fitting the modern brewer’s needs. Cerevisia 26, 89–97 Genetics 154, 99–107
62 De Vit, M.J. et al. (1997) Regulated nuclear translocation of the Mig1
52 Van Dijck, P. et al. (2000) A baker’s yeast mutant (fil1) with a specific,
glucose repressor. Mol. Biol. Cell 8, 1603–1618
partially inactivating mutation in adenylate cyclase maintains a high
63 Östling, J. and Ronne, H. (1998) Negative control of the Mig1
stress resistance during active fermentation and growth. J. Mol.
represssor by Snf1-dependent phosphorylation in the absence of
Microbiol. Biotechnol. 2, 521–530
glucose. Eur. J. Biochem. 252, 162–168
53 Patnaik, R. et al. (2002) Genome shuffling of Lactobacillus for
64 Colombo, S. et al. (1998) Involvement of distinct G-proteins Gpa2 and
improved acid tolerance. Nat. Biotechnol. 20, 707–712
Ras, in glucose- and intracellular acidification-induced cAMP signal-
54 Boles, E. et al. Recombinant Saccharomyces cerevisiae expressing
ling in the yeast Saccharomyces cerevisiae. EMBO J. 17, 3326–3341
chimeric glucose transporters. Patent WO200880. Gothia Yeast
65 Toda, T. et al. (1987) Cloning and characterization of Bcy1, a locus
Solutions AB (2002) encoding a regulatory subunit of the cyclic AMP-dependent protein-
55 Park, J.I. et al. (1997) The freeze–thaw stress response of the yeast kinase in Saccharomyces cerevisiae. Mol. Cell. Biol. 7, 1371–1377
Saccharomyces cerevisiae is growth phase specific and is controlled by 66 Toda, T. et al. (1987) 3 different genes in Saccharomyces cerevisiae
nutritional state via the RAS-cyclic AMP signal transduction path- encode the catalytic subunits of the cAMP-dependent protein kinase.
way. Appl. Environ. Microbiol. 63, 3818–3824 Cell 50, 277–287
56 Dumortier, F. et al. (1999) New strains ‘fil’’, stress-resistant under 67 Broach, J. et al. (1985) Ras proteins function exclusively to modulate
fermentation and/or growth conditions. Patent EP0967280. Lesaffre & adenylate cyclase activity in the yeast Saccharomyces DNA. J. Mol.
Cie, France Cell. Biol. 4, 64
57 Kraakman, L. et al. (1999) A Saccharomyces cerevisiae G-protein 68 Toda, T. et al. (1985) In yeast, Ras proteins are controlling elements of
coupled receptor Gpr1, is specifically required for glucose activation of adenylate cyclase. Cell 40, 27–36
the cAMP pathway during the transition to growth on glucose. Mol. 69 Griffioen, G. and Thevelein, J.M. (2002) Molecular mechanisms
Microbiol. 32, 1002–1012 controlling the localisation of protein kinase A. Curr. Genet. 41, 199–207

ScienceDirect collection reaches six million full-text articles

Elsevier recently announced that six million articles are now available on its premier electronic platform, ScienceDirect. This milestone
in electronic scientific, technical and medical publishing means that researchers around the globe will be able to access an unsurpassed
volume of information from the convenience of their desktop.
The rapid growth of the ScienceDirect collection is due to the integration of several prestigious publications as well as ongoing addition
to the Backfiles - heritage collections in a number of disciplines. The latest step in this ambitious project to digitize all of Elsevier’s
journals back to volume one, issue one, is the addition of the highly cited Cell Press journal collection on ScienceDirect. Also available
online for the first time are six Cell titles’ long-awaited Backfiles, containing more than 12,000 articles highlighting important historic
developments in the field of life sciences.

www.sciencedirect.com

www.sciencedirect.com